Selective [1,2,4]Triazolo[4,3-a] Heterocycles as Anticancer Agents: In Vitro and In Silico Analysis

In this study, 1,2,4-triazole fused heterocycles such as [1,2,4]triazolo[4,3-a]pyridine, [1,2,4]triazolo[4,3-a]quinoline, and [1,2,4]triazolo[4,3-a]pyrimidine derivatives using substituted benzylamine and 2-hydrazino heterocyclic ring under ambient reaction condition were designed and synthesized. These synthesized compounds were subsequently evaluated for their biological activities, with a focus on anticancer potential. Initial in vitro studies revealed promising results in the inhibition of growth in breast and colon cancer cell lines. The most potent compound 10, was further analyzed for cancer cell-killing potential using fluorescence imaging and TEM. Further, the wound healing assay on breast cancer cell lines (KAIMRC2) and the human colon cancer cell line (HCT8) was performed by treating compound 10, and it was observed that HCT8 cell line exhibited faster wound closure compared with KAIMRC2. The Western blot analysis was performed on cancer cells to assess efficacy in modulating key signaling pathways in apoptosis. These results provide strong evidence that compound 10 induces apoptotic cell death through unknown pathways and may serve as a promising anticancer agent. Furthermore, to rationalize the observed biological activity, the molecular docking and dynamic simulations have also been carried out to understand the binding affinity and binding interactions of enzymes and synthesized derivatives. This revealed a significant correlation between these compounds’ binding score and biological activity. The results of the in vitro and in silico studies suggest that the [1,2,4]triazolo[4,3-a]pyridine scaffold could be the new stepping stone to achieve anticancer agents.